Crystalline form of benzylbenzene sglt2 inhibitor

ABSTRACT

Provided are crystalline forms of a compound having an inhibitory effect on sodium-dependent glucose cotransporter SGLT2. The invention also provides pharmaceutical compositions, methods of preparing the crystalline compound, and methods of using the crystalline compound, independently or in combination with other therapeutic agents, for treating diseases and conditions which are affected by SGLT or SGLT2 inhibition.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority to PCT/CN2010/073865, filed Jun. 12,2010, which is incorporated in its entirety herein for all purposes.

BACKGROUND OF THE INVENTION

The sodium-dependent (“active”) glucose cotransporters (SGLTs),including SGLT1 (found predominantly in the intestinal brush border) andSGLT2 (localized in the renal proximal tubule), have been significantlyevaluated. In particular, SGLT2 has been found to be responsible for themajority of glucose reuptake by the kidneys. Inhibition of renal SGLT isnow considered a useful approach to treating hyperglycemia by increasingthe amount of glucose excreted in the urine (Arakawa K, et al., Br JPharmacol 132:578-86, 2001; Oku A, et al., Diabetes 48:1794-1800, 1999).The potential of this therapeutic approach is further supported byrecent findings that mutations in the SGLT2 gene occur in cases offamilial renal glucosuria, an apparently benign syndrome characterizedby urinary glucose excretion in the presence of normal serum glucoselevels and the absence of general renal dysfunction or other disease(Santer R, et al., J Am Soc Nephrol 14:2873-82, 2003). Therefore,compounds which inhibit SGLT, particularly SGLT2, are promisingcandidates for use as antidiabetic drugs (reviewed in Washburn W N,Expert Opin Ther Patents 19:1485-99, 2009). In addition, since cancercells show increased glucose uptake in comparison to their normalcounterparts, SGLT inhibition has been proposed as a method for treatingcancer by starving cancer cells. For example, studies suggest that SGLT2plays a role in glucose uptake in metastatic lesions of lung cancer(Ishikawa N, et al., Jpn J Cancer Res 92:874-9, 2001). Thus, SGLT2inhibitors may also be useful as anticancer agents.

In addition to pharmaceutical activity, a further consideration for thesuccessful development of a medicament is the parameters which areconnected with the physical nature of the active substance itself. Someof these parameters are stability of the active substance under variousenvironmental conditions, stability of the active substance duringproduction of the pharmaceutical formulation and the stability of theactive substance in the final medicament compositions. In order toprovide the necessary stability, the pharmaceutically active substanceused in the medicament should be as pure as possible, leading to itsstability in long-term storage under various environmental conditions.

Another factor to be considered is the uniform distribution of theactive substance in the formulation, particularly when the activesubstance is to be given in low doses. To ensure uniform distribution,the particle size of the active substance can be reduced to a suitablelevel, e.g. by grinding. However, breakdown of the pharmaceuticallyactive substance as a side effect of the grinding (or micronising) mustbe avoided. As a result, in view of the hard conditions required duringthe process, the active substance must be stable throughout the grindingprocess. Still further, if the active substance is not stable during thegrinding process, a homogeneous pharmaceutical formulation with thespecified amount of active substance is unlikely to be achieved in areproducible manner.

Still another consideration associated with the grinding process forpreparing the desired pharmaceutical formulation is the input of energycaused by this process and the stress on the surface of the crystals.This may in certain circumstances lead to polymorphous changes, toamorphization or to a change in the crystal lattice. Since thepharmaceutical quality of a pharmaceutical formulation requires that theactive substance should always have the same crystalline morphology, thestability and properties of the crystalline active substance are subjectto stringent requirements from this point of view as well.

Another consideration for the pharmaceutically active substance isstability in a formulation, which in turn gives rise to a longer shelflife of the particular medicament. In this instance, the shelf life isthe length of time during which the medicament can be administeredwithout any risk that the active substance has degraded. High stabilityof a medicament in the abovementioned pharmaceutical compositions undervarious storage conditions is therefore an additional advantage for boththe patient and the manufacturer.

Furthermore, the availability of a well-defined crystalline form allowsthe purification of the drug substance by recrystallization.

Apart from the requirements indicated above, it should be generallyborne in mind that any change to the solid state of a pharmaceuticalcomposition which is capable of improving its physical and chemicalstability gives a significant advantage over less stable forms of thesame medicament.

The compound of the present invention has been prepared according to themethods of U.S. Publication No. 2009/0118201, filed Aug. 22, 2008, U.S.application Ser. No. 12/545,400, and PCT/US2009/054585, nowWO2010/022313, both filed Aug. 21, 2009. The aim of the presentinvention is to provide a stable crystalline form of the compound whichmeets important requirements imposed on pharmaceutically activesubstances as mentioned above.

BRIEF SUMMARY OF THE INVENTION

The present invention provides crystalline forms of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolhaving an inhibitory effect on sodium-dependent glucose cotransporter 2(SGLT2). The invention also provides pharmaceutical compositions,methods of preparing the crystalline form of the compound, and methodsof using the compound, independently or in combination with othertherapeutic agents, for treating diseases and conditions which areaffected by SGLT2 inhibition.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides the X-ray powder diffraction (XRPD) spectra ofcrystalline(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

FIG. 2 provides a Table of XRPD data for the XRPD spectra in FIG. 1.

FIG. 3 provides the Raman spectra of crystalline(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

FIG. 4 provides a Raman peak list for the Raman spectra in FIG. 3.

FIG. 5 provides the thermal gravimetric analysis (TGA) of crystalline(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

FIG. 6 provides the differential scanning calorimetry (DSC) spectra ofcrystalline(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

FIG. 7 provides a table of unit cell data for crystalline(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

FIG. 8 provides a scheme for the preparation of crystalline(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

DETAILED DESCRIPTION OF THE INVENTION I. Definitions

As used herein, the terms “treating” and “treatment” refer to delayingthe onset of, retarding or reversing the progress of, or alleviating orpreventing either the disease or condition to which the term applies, orone or more symptoms of such disease or condition.

As used herein, the term “administering” means oral administration,administration as a suppository, topical contact, intravenous,intraperitoneal, intramuscular, intralesional, intranasal orsubcutaneous administration, or the implantation of a slow-releasedevice, e.g., a mini-osmotic pump, to a subject. Administration is byany route including parenteral, and transmucosal (e.g., oral, nasal,vaginal, rectal, or transdermal). Parenteral administration includes,e.g., intravenous, intramuscular, intra-arteriole, intradermal,subcutaneous, intraperitoneal, intraventricular, and intracranial. Othermodes of delivery include, but are not limited to, the use of liposomalformulations, intravenous infusion, transdermal patches, and the like.

As used herein, the term “prodrug” refers to a precursor compound that,following administration, releases the biologically active compound invivo via some chemical or physiological process (e.g., a prodrug onreaching physiological pH or through enzyme action is converted to thebiologically active compound). A prodrug itself may either lack orpossess the desired biological activity.

As used herein, the term “compound” refers to a molecule produced by anymeans including, without limitation, synthesis in vitro or generation insitu or in vivo.

The terms “controlled release,” “sustained release,” “extended release,”and “timed release” are intended to refer interchangeably to anydrug-containing formulation in which release of the drug is notimmediate, i.e., with a “controlled release” formulation, oraladministration does not result in immediate release of the drug into anabsorption pool. The terms are used interchangeably with “nonimmediaterelease” as defined in Remington: The Science and Practice of Pharmacy,21^(St) Ed., Gennaro, Ed., Lippencott Williams & Wilkins (2003). Asdiscussed therein, immediate and nonimmediate release can be definedkinetically by reference to the following equation:

The “absorption pool” represents a solution of the drug administered ata particular absorption site, and k_(r), k_(a) and k_(e) are first-orderrate constants for (1) release of the drug from the formulation, (2)absorption, and (3) elimination, respectively. For immediate releasedosage forms, the rate constant for drug release k_(r) is far greaterthan the absorption rate constant k_(a). For controlled releaseformulations, the opposite is true, i.e., k_(r)<<k_(a), such that therate of release of drug from the dosage form is the rate-limiting stepin the delivery of the drug to the target area.

The terms “sustained release” and “extended release” are used in theirconventional sense to refer to a drug formulation that provides forgradual release of a drug over an extended period of time, for example,12 hours or more, and that preferably, although not necessarily, resultsin substantially constant blood levels of a drug over an extended timeperiod.

As used herein, the term “delayed release” refers to a pharmaceuticalpreparation that passes through the stomach intact and dissolves in thesmall intestine.

As used herein, the term “pharmaceutically acceptable excipient” refersto a substance that aids the administration of an active agent to andabsorption by a subject. Pharmaceutical excipients useful in the presentinvention include, but are not limited to, binders, fillers,disintegrants, lubricants, coatings, sweeteners, flavors and colors. Oneof skill in the art will recognize that other pharmaceutical excipientsare useful in the present invention.

As used herein, the term “subject” refers to animals such as mammals,including, but not limited to, primates (e.g., humans), cows, sheep,goats, horses, dogs, cats, rabbits, rats, mice and the like. In certainembodiments, the subject is a human.

As used herein, the terms “therapeutically effective amount or dose” or“therapeutically sufficient amount or dose” or “effective or sufficientamount or dose” refer to a dose that produces therapeutic effects forwhich it is administered. The exact dose will depend on the purpose ofthe treatment, and will be ascertainable by one skilled in the art usingknown techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms(vols. 1-3, 1992); Lloyd, The Art, Science and Technology ofPharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999);and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003,Gennaro, Ed., Lippincott, Williams & Wilkins).

II. Crystalline Forms

The present invention provides a crystalline form of a compound havingan inhibitory effect on sodium-dependent glucose cotransporter SGLT,preferably SGLT2. Therefore, the crystalline compound of the presentinvention is suitable for the prevention and treatment of diseases andconditions, particularly metabolic disorders, including but not limitedto type 1 and type 2 diabetes mellitus, hyperglycemia, diabeticcomplications (such as retinopathy, nephropathy, e.g., progressive renaldisease, neuropathy, ulcers, micro- and macroangiopathies, and diabeticfoot disease), insulin resistance, metabolic syndrome (Syndrome X),hyperinsulinemia, hypertension, hyperuricemia, obesity, edema,dyslipidemia, chronic heart failure, atherosclerosis and relateddiseases.

The present invention also provides pharmaceutical compositions andprodrugs of the crystalline form according to the present invention.

The present invention further provides synthetic processes for preparingthe crystalline compound of the present invention.

The present invention also provides methods of using the crystallineform of the compound according to the present invention, independentlyor in combination with other therapeutic agents, for treating diseasesand conditions which may be affected by SGLT inhibition.

The present invention also provides methods of using the compoundsaccording to the present invention for the preparation of a medicamentfor treating diseases and conditions which may be affected by SGLTinhibition.

In one aspect, the present invention provides a crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.The chemical structure is shown below:

The crystalline compound of the present invention can be characterizedby the X-ray powder diffraction (XRPD), the Raman spectra, thedifferential scanning calorimetry (DSC) endotherm, the thermalgravimetric analysis (TGA) showing decomposition temperature, and theunit cell of the crystal structure.

In some embodiments, the present invention provides the crystalline formof the compound characterized by the XRPD substantially in accordancewith that of FIG. 1 and the peaks substantially in accordance with FIG.2. The crystalline compound of the present invention can have anycombination of peaks substantially in accordance with FIG. 2. Moreover,each peak listed in FIG. 2 can have an error range of ±0.2 degrees 2θ,preferably ±0.1 degrees 2θ.

In other embodiments, the crystalline form of the compound ischaracterized by an X-ray powder diffraction pattern that includes oneor more peaks at 5.4, 11.2, 11.3, 11.9, 12.9, 15.5, 16.3, 17.8, 19.1,20.0, 20.6, 20.7, 21.2, 22.8, 23.0, 23.4, 23.6, 23.9, 24.7, 25.4, 25.8,27.8 and 28.2 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is madeusing CuK_(α1) radiation. In another embodiment, the crystalline form ofthe compound is characterized by an XRPD that includes two or more,three or more, four or more, or five or more peaks at 5.4, 11.2, 11.3,11.9, 12.9, 15.5, 16.3, 17.8, 19.1, 20.0, 20.6, 20.7, 21.2, 22.8, 23.0,23.4, 23.6, 23.9, 24.7, 25.4, 25.8, 27.8 and 28.2 degrees 2θ (±0.1degrees 2θ). In some other embodiments, the crystalline form of thecompound is characterized by an XRPD that includes peaks at 12.9, 19.1and 20.7 degrees 2θ (±0.1 degrees 2θ). In still other embodiments, thecrystalline form of the compound is characterized by an XRPD thatincludes peaks at 11.2, 12.9, 15.5, 17.8, 19.1, 20.0 and 20.7 degrees 2θ(±0.1 degrees 2θ). In yet other embodiments, the crystalline form of thecompound is characterized by an XRPD that includes peaks at 5.4, 11.2,11.9, 12.9, 15.5, 16.3, 17.8, and 19.1 degrees 2θ (±0.1 degrees 2θ). Instill yet other embodiments, the crystalline form of the compound ischaracterized by an XRPD that includes peaks at 5.4, 11.2, 11.9, and12.9 degrees 2θ (±0.1 degrees 2θ). In another embodiment, thecrystalline form of the compound is characterized by an XRPD includingpeaks at 11.2 and 12.9 degrees 2θ (±0.1 degrees 2θ). In otherembodiments, the crystalline form of the compound is characterized bythe XRPD peaks substantially in accordance with FIG. 2.

The crystalline compound of the present invention is also characterizedby the Raman spectra substantially in accordance with FIG. 3 and thepeaks substantially in accordance with FIG. 4. In some embodiments, thecrystalline form of the compound is characterized by a Raman spectrathat includes one or more peaks at about 353, 688, 825, 1178, 1205,1212, 1608, 2945, 3010 and 3063 cm⁻¹. In another embodiment, thecrystalline form of the compound is characterized by a Raman spectrathat includes two or more, three or more, four or more, or five or morepeaks. In other embodiments, the crystalline form of the compound ischaracterized by the Raman spectra including peaks at about 353, 688 and825 cm⁻¹. In some other embodiments, the crystalline form of thecompound is characterized by the Raman peaks substantially in accordancewith FIG. 4.

The crystalline compound of the present invention is also characterizedby the differential scanning calorimetry (DSC) endotherm. In someembodiments, the crystalline form of the compound is characterized by aDSC endotherm at about 136° C.

The crystalline compound of the present invention is also characterizedby the unit cell data substantially in accordance with FIG. 7. Thermalgravimetric analysis (TGA) can also be used to characterize thecrystalline compound of the present invention. For example, arepresentative TGA is substantially in accordance with that shown inFIG. 5, demonstrating thermal stability of the crystalline compoundabove 200° C.

In some embodiments, the crystalline compound is characterized by atleast one of the following: at least one XRPD peak as described above,at least one Raman peak as described above, a DSC endotherm as describedabove, TGA data regarding thermal stability as described above, and unitcell data as described above and in FIG. 7. In other embodiments, thecrystalline compound is characterized by at least two of the following:at least one XRPD peak as described above, at least one Raman peak asdescribed above, a DSC endotherm as described above, TGA data regardingthermal stability as described above, and unit cell data as describedabove and in FIG. 7. For example, the crystalline compound can becharacterized by at least one XRPD peak and at least one Raman peak, orat least one XRPD peak and the DSC endotherm, or at least one Raman peakand the DSC endotherm, or at least one XRPD peak and the unit cell data,or at least one Raman peak and the unit cell data, etc.

In some embodiments, the crystalline compound of the present inventionis characterized by an X-ray powder diffraction (XRPD) pattern thatincludes one or more peaks at 5.4, 11.2, 11.3, 11.9, 12.9, 15.5, 16.3,17.8, 19.1, 20.0, 20.6, 20.7, 21.2, 22.8, 23.0, 23.4, 23.6, 23.9, 24.7,25.4, 25.8, 27.8 and 28.2 degrees 2θ (±0.1 degrees 2θ), wherein saidXRPD is made using CuK_(α1) radiation, and a Raman spectra that includesone or more peaks at about 353, 688, 825, 1178, 1205, 1212, 1608, 2945,3010 and 3063 cm⁻¹. In other embodiments, the crystalline compound ofthe present invention is characterized by an X-ray powder diffraction(XRPD) pattern that includes one or more peaks at 11.2, 12.9, 15.5,17.8, 19.1, 20.0, 20.6, 20.7, 21.2 and 22.8 and 28.2 degrees 2θ (±0.1degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation, and aRaman spectra that includes one or more peaks at about 353, 688, and 825cm⁻¹. In some other embodiments, the crystalline compound of the presentinvention is characterized by an X-ray powder diffraction (XRPD) patternthat includes one or more peaks at 11.2 and 12.9 degrees 2θ (±0.1degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation, and aRaman spectra that includes one or more peaks at about 353, 688, and 825cm⁻¹.

In other embodiments, the present invention provides a compound(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolin crystalline form.

The present invention also includes isotopically-labeled forms of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,wherein one or more atoms are replaced by one or more atoms havingspecific atomic mass or mass numbers. Examples of isotopes that can beincorporated into compounds of the invention include, but are notlimited to, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine,sulfur, and chlorine (such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ¹⁸F, ³⁵Sand ³⁶Cl). Isotopically-labeled compounds and prodrugs thereof, as wellas isotopically-labeled, pharmaceutically acceptable salts and prodrugsthereof, are within the scope of the present invention.Isotopically-labeled compounds of the present invention are useful inassays of the tissue distribution of the compounds and their prodrugsand metabolites; preferred isotopes for such assays include ³H and ¹⁴C.In addition, in certain circumstances substitution with heavierisotopes, such as deuterium (²H), can provide increased metabolicstability, which offers therapeutic advantages such as increased in vivohalf-life or reduced dosage requirements. Isotopically-labeled compoundsof this invention and prodrugs thereof can generally be preparedaccording to the methods described herein by substituting anisotopically-labeled reagent for a non-isotopically labeled reagent.

III. Methods of Making Crystalline Form

The crystalline form of the present invention can be obtained by avariety of methods, as outlined in FIG. 8. For example, the crystallinecompound 8 can be prepared directly from L-proline complex 7.Alternatively, the L-proline of L-proline complex 7 can be removed toafford amorphous 8, which is then crystallized to crystalline 8.Crystalline 8 can also be prepared directly from crude compound 6, byfirst isolating and then crystallizing amorphous 8 to form crystalline8, or directly from crude 6.

Other methods of preparing crystalline 8 are know to one of skill in theart. Moreover, each crystallization process can be repeated to removeadditional impurities. In some embodiments, more than one of the variouscrystallization processes can be used to prepare the crystallinecompound of the present invention.

In some embodiments, crystalline 8 can be prepared from the bisL-proline co-crystal of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolas described in the Examples. Briefly, the co-crystal starting materialsis taken up in a suitable solvent (e.g., methanol or ethanol) to obtaina solution, and a precipitating solvent (e.g., water) is added toachieve crystallization of the desired compound.

Accordingly, the present invention further provides a method for makinga crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,the method including (a) combining(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolbis(L-proline) complex and a suitable solvent with mixing to form asolution; (b) adding a precipitating solvent to the solution to providea mixture; and (c) isolating the crystalline form from the mixture ofstep (b).

In some embodiments, the present invention provides a method for makinga crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,the method including (a) combining(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-trioland a suitable solvent with mixing to form a solution; (b) adding aprecipitating solvent to the solution to provide a mixture; and (c)isolating the crystalline form from the mixture of step (b).

In other embodiments, the present invention provides a method for makinga crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,the method including (a) combining amorphous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-trioland a suitable solvent with mixing to form a solution; (b) adding aprecipitating solvent to the solution to provide a mixture; and (c)isolating the crystalline form from the mixture of step (b).

In step (a) of the above methods, the solvent can be any solventsuitable to form a solution, and which is miscible with theprecipitating solvent used in step (b). Typically the solvent in step(a) is a polar solvent, which in some embodiments is a protic solvent.Suitable solvents include, C₁-C₄ alcohols, ethylene glycol andpolyethylene glycol such as PEG400, alkanoates such as ethyl acetate,isopropyl acetate, propyl acetate, and butyl acetate, acetonitrile,alkanones such as acetone, butanone, methyl ethyl ketone and methylpropyl ketone, and mixtures of two or more of these solvents. Morepreferred solvents are selected from the group consisting of methanol,ethanol, isopropanol, ethyl acetate, acetone, and mixture of two or moreof these solvents. Still further preferred are methanol and ethanol. Inone selected embodiment, the solvent used in step (a) is methanol.

Step (a) of the above methods can be carried out at temperaturesgenerally from about 0° C. to the reflux temperature of the solvent(e.g., 65° C. for methanol). A preferred temperature range is betweenabout 35° C. and 100° C., even more preferably from about 45° C. to 80°C. Once a solution is obtained, a precipitating solvent is added. Aprecipitating solvent is one in which the product is much less solublethan the initial solution solvent. Suitable precipitating solventsinclude water, ethers, cyclic ethers, alkanes, cycloalkanes, phenyls andmixtures thereof, in particular C₄-C₆-aliphatic ethers, C₆-C₈-alkanes,C₆-C₈-cycloalkanes, phenyls such as benzene, toluene and xylene, andmixtures thereof. Examples of precipitating solvents arediisopropylether, tert-butylmethylether (TBME), cyclohexane,methylcyclohexane, hexane, heptane, octane and mixtures thereof. In oneselected embodiment, the precipitating solvent is water.

The precise ratios of solvents and starting material are less criticalto the invention, but optimized ratios can produce greater yields andmore uniform crystallized product. The ratio of solvents in the abovemethods can be any suitable ratio from about 1:1 to about 1:9, includingabout 1:2, 1:3, 1:4, 1:5, 1:6, 1:7 and about 1:8. The range of solventratios is preferably from about 1:1 to about 1:9, more preferably fromabout 1:2 to about 1:7, even more preferably from about 1:2 to about1:5. In one group of embodiments, when methanol is used as solvent andwater is the precipitating solvent, the ratio of methanol to water inthe mixture of step (b) is from about 1:1 to about 1:9 by volume, morepreferably about 1:5 by volume.

The ratio of complex to solvent, such as a methanol and water mixture,can be any suitable ratio to promote crystallization. For example, thecomplex to solvent ratio can be from about 1:5 (weight/volume, or w/v)to about 1:50 (w/v), including about 1:6, 1:7, 1:8, 1:9, 1:10, 1:11,1:12, 1:13, 1:14, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40 and about 1:45(w/v). The complex to solvent ratio is preferably from about 1:10 toabout 1:25 (w/v), more preferably from about 1:10 to about 1:15 (w/v).In another group of embodiments, the ratio of complex to solvent andprecipitating solvent in the mixture of step (b) is from about 1:10 toabout 1:25 (w/v). In other embodiments, the ratio of complex to methanoland water in the mixture of step (b) is from about 1:10 to about 1:25(w/v). In some other embodiments, the ratio of complex to methanol andwater in the mixture of step (b) is from about 1:2:7 (w/v/v) to about1:3:10 (w/v/v), preferably about 1:2:10 (w/v/v).

The mixture for crystallizing crystalline 8 can also contain a varietyof other components, such as acids, bases and salts. Acids useful in thepresent invention include, but are not limited to, acetic acid, formicacid, hydrochloric acid, sulfuric acid, and other weak acids and strongacids. Bases useful in the present invention include, but are notlimited to, ammonia, sodium hydroxide, and others. Salts useful in thepresent invention include, but are not limited to, sodium chloride,potassium chloride, potassium carbonate and others. In some embodiments,the mixture of step (b) in the above methods includes sodium hydroxide.In other embodiments, the mixture of step (b) in the above methodsincludes sodium chloride.

After addition of the precipitating solvent, the mixture is generallykept at room temperature, or cooled, for a sufficient period of time toallow complete crystal formation of the product to occur. Thetemperature of the mixture in step (b) is preferably about the same asor lower than in step (a). During storage the temperature of thesolution containing the product is preferably lowered to a temperaturein the range from −10° C. to 25° C. or even lower, even more preferablyin the range from −5° C. to 15° C. Step (b) can be carried out with orwithout stirring. As noted above, the conditions for step (b) can affectthe size, shape and quality of the obtained crystals.

Crystallization can be induced by methods known in the art, for exampleby mechanical means such as scratching or rubbing the contact surface ofthe reaction vessel with e.g. a glass rod. Optionally the saturated orsupersaturated solution may be inoculated with seed crystals. Themixture for crystallizing crystalline 8 can also contain a seed crystalof crystalline compound 8. In some embodiments, the solution or mixturein the above methods includes a seed crystal of the crystalline compoundof the present invention.

Isolation of the desired crystalline form can be accomplished byremoving the solvent and precipitating solvent from the crystals.Generally this is carried out by known methods as for examplefiltration, suction filtration, decantation or centrifugation. Furtherisolation can be achieved by removing any excess of the solvent(s) fromthe crystalline form by methods known to the one skilled in the art asfor example application of a vacuum, and/or by heating above 20° C.,preferably in a temperature range below 80° C., even more preferablybelow 50° C.

In other embodiments, the present invention provides a method for makinga crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,the method including (a) combining(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-trioland a suitable solvent with mixing to form a solution; and (b) isolatingthe crystalline form from the solution. In other embodiments, the methodalso includes adding a precipitating solvent to the solution. In step(a) of the above methods, the solvent can be any solvent suitable toform a solution. Suitable solvents include alkanoates such as ethylacetate, isopropyl acetate, propyl acetate, and butyl acetate, etherssuch as ethyl ether, methyl tert-butyl ether and mixtures of two or moreof these solvents. More preferred solvents are selected from the groupconsisting of ethyl acetate, ethyl ether, methyl tert-butyl ether andmixture of two or more of these solvents. Still further preferred areethyl acetate and methyl tert-butyl ether. The(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolcan have any suitable form, including amorphous, crystalline, or acombination thereof. Moreover, the(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolcan have any suitable level of purity, such as purified or unpurified.

In some embodiments, the(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolis amorphous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.The amorphous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,amorphous compound 8, can be prepared by a variety of methods known inthe art. For example, the amorphous compound 8 can be isolated fromcrude mixture 6 using known methods of isolating. Alternatively,amorphous compound 8 can be prepared from complex 7 by removing theL-proline using methods known in the art. In some embodiments, theamorphous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolis prepared from(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolbis(L-proline) by combining(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolbis(L-proline) complex and a suitable solvent mixture with mixing toform a solution, and isolating amorphous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolfrom the solution. Suitable solvents and solvent mixtures are describedabove.

IV. Pharmaceutical Compositions

The present invention further provides a pharmaceutical compositioncomprising an effective amount of a crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,in a pharmaceutically acceptable excipient.

The crystalline form provided in this invention can be incorporated intoa variety of formulations for therapeutic administration. Moreparticularly, the crystalline form of the present invention can beformulated into pharmaceutical compositions, by formulation withappropriate pharmaceutically acceptable excipients or diluents, and canbe formulated into preparations in solid, semi-solid, liquid or gaseousforms, such as tablets, capsules, pills, powders, granules, dragees,gels, slurries, ointments, solutions, suppositories, injections,inhalants and aerosols. As such, administration of the crystalline formof the present invention can be achieved in various ways, includingoral, buccal, parenteral, intravenous, intradermal (e.g., subcutaneous,intramuscular), transdermal, etc., administration. Moreover, thecrystalline form can be administered in a local rather than systemicmanner, for example, in a depot or sustained release formulation.

Suitable formulations for use in the present invention are found inRemington: The Science and Practice of Pharmacy, 21^(st) Ed., Gennaro,Ed., Lippencott Williams & Wilkins (2003), which is hereby incorporatedherein by reference. The pharmaceutical compositions described hereincan be manufactured in a manner that is known to those of skill in theart, i.e., by means of conventional mixing, dissolving, granulating,dragee-making, levigating, emulsifying, encapsulating, entrapping orlyophilizing processes. The following methods and excipients are merelyexemplary and are in no way limiting.

In one preferred embodiment, the crystalline form of the presentinvention is prepared for delivery in a sustained-release, controlledrelease, extended-release, timed-release or delayed-release formulation,for example, in semipermeable matrices of solid hydrophobic polymerscontaining the therapeutic agent. Various types of sustained-releasematerials have been established and are well known by those skilled inthe art. Current extended-release formulations include film-coatedtablets, multiparticulate or pellet systems, matrix technologies usinghydrophilic or lipophilic materials and wax-based tablets withpore-forming excipients (see, for example, Huang, et al. Drug Dev. Ind.Pharm. 29:79 (2003); Pearnchob, et al. Drug Dev. Ind. Pharm. 29:925(2003); Maggi, et al. Eur. J. Pharm. Biopharm. 55:99 (2003); Khanvilkar,et al., Drug Dev. Ind. Pharm. 228:601 (2002); and Schmidt, et al., Int.J. Pharm. 216:9 (2001)). Sustained-release delivery systems can,depending on their design, release the compounds over the course ofhours or days, for instance, over 4, 6, 8, 10, 12, 16, 20, 24 hours ormore. Usually, sustained release formulations can be prepared usingnaturally-occurring or synthetic polymers, for instance, polymeric vinylpyrrolidones, such as polyvinyl pyrrolidone (PVP); carboxyvinylhydrophilic polymers; hydrophobic and/or hydrophilic hydrocolloids, suchas methylcellulose, ethylcellulose, hydroxypropylcellulose, andhydroxypropylmethylcellulose; and carboxypolymethylene.

The sustained or extended-release formulations can also be preparedusing natural ingredients, such as minerals, including titanium dioxide,silicon dioxide, zinc oxide, and clay (see, U.S. Pat. No. 6,638,521,herein incorporated by reference). Exemplified extended releaseformulations that can be used in delivering a compound of the presentinvention include those described in U.S. Pat. Nos. 6,635,680;6,624,200; 6,613,361; 6,613,358, 6,596,308; 6,589,563; 6,562,375;6,548,084; 6,541,020; 6,537,579; 6,528,080 and 6,524,621, each of whichis hereby incorporated herein by reference. Controlled releaseformulations of particular interest include those described in U.S. Pat.Nos. 6,607,751; 6,599,529; 6,569,463; 6,565,883; 6,482,440; 6,403,597;6,319,919; 6,150,354; 6,080,736; 5,672,356; 5,472,704; 5,445,829;5,312,817 and 5,296,483, each of which is hereby incorporated herein byreference. Those skilled in the art will readily recognize otherapplicable sustained release formulations.

For oral administration, the crystalline form of the present inventioncan be formulated readily by combining with pharmaceutically acceptableexcipients that are well known in the art. Such excipients enable thecompounds to be formulated as tablets, pills, dragees, capsules,emulsions, lipophilic and hydrophilic suspensions, liquids, gels,syrups, slurries, suspensions and the like, for oral ingestion by apatient to be treated. Pharmaceutical preparations for oral use can beobtained by mixing the compounds with a solid excipient, optionallygrinding a resulting mixture, and processing the mixture of granules,after adding suitable auxiliaries, if desired, to obtain tablets ordragee cores. Suitable excipients are, in particular, fillers such assugars, including lactose, sucrose, mannitol, or sorbitol; cellulosepreparations such as, for example, maize starch, wheat starch, ricestarch, potato starch, gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/orpolyvinylpyrrolidone (PVP). If desired, disintegrating agents can beadded, such as a cross-linked polyvinyl pyrrolidone, agar, or alginicacid or a salt thereof such as sodium alginate.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds can be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers can be added. All formulations fororal administration should be in dosages suitable for suchadministration.

Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions can be used, which can optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments can be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

The crystalline forms described herein can also be formulated forparenteral administration by injection, e.g., by bolus injection orcontinuous infusion. For injection, the compound can be formulated intopreparations by dissolving, suspending or emulsifying them in an aqueousor nonaqueous solvent, such as vegetable or other similar oils,synthetic aliphatic acid glycerides, esters of higher aliphatic acids orpropylene glycol; and if desired, with conventional additives such assolubilizers, isotonic agents, suspending agents, emulsifying agents,stabilizers and preservatives. In some embodiments, the crystallineforms of the invention can be formulated in aqueous solutions,preferably in physiologically compatible buffers such as Hank'ssolution, Ringer's solution, or physiological saline buffer.Formulations for injection can be presented in unit dosage form, e.g.,in ampoules or in multi-dose containers, with an added preservative. Thecompositions can take such forms as suspensions, solutions or emulsionsin oily or aqueous vehicles, and can contain formulatory agents such assuspending, stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration includeaqueous solutions of the active compounds in water-soluble form.Additionally, suspensions of the active compounds can be prepared asappropriate oily injection suspensions. Suitable lipophilic solvents orvehicles include fatty oils such as sesame oil, or synthetic fatty acidesters, such as ethyl oleate or triglycerides, or liposomes. Aqueousinjection suspensions can contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension can also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.Alternatively, the active ingredient can be in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

Systemic administration can also be by transmucosal or transdermalmeans. For transmucosal or transdermal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.For topical administration, the agents are formulated into ointments,creams, salves, powders and gels. In one embodiment, the transdermaldelivery agent can be DMSO. Transdermal delivery systems can include,e.g., patches. For transmucosal administration, penetrants appropriateto the barrier to be permeated are used in the formulation. Suchpenetrants are generally known in the art. Exemplified transdermaldelivery formulations that can find use in the present invention includethose described in U.S. Pat. Nos. 6,589,549; 6,544,548; 6,517,864;6,512,010; 6,465,006; 6,379,696; 6,312,717 and 6,310,177, each of whichare hereby incorporated herein by reference.

For buccal administration, the compositions can take the form of tabletsor lozenges formulated in conventional manner.

In addition to the formulations described previously, a crystalline formof the present invention can also be formulated as a depot preparation.Such long acting formulations can be administered by implantation (forexample subcutaneously or intramuscularly) or by intramuscularinjection. Thus, for example, the compounds can be formulated withsuitable polymeric or hydrophobic materials (for example as an emulsionin an acceptable oil) or ion exchange resins, or as sparingly solublederivatives, for example, as a sparingly soluble salt.

The pharmaceutical compositions also can comprise suitable solid or gelphase carriers or excipients. Examples of such carriers or excipientsinclude but are not limited to calcium carbonate, calcium phosphate,various sugars, starches, cellulose derivatives, gelatin, and polymerssuch as polyethylene glycols.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions wherein the active ingredients are contained in atherapeutically effective amount. The present invention alsocontemplates pharmaceutical compositions comprising the crystallineforms of the invention in admixture with an effective amount of othertherapeutic agents as combination partners, particularly those used fortreating diseases and conditions which can be affected by SGLTinhibition, such as antidiabetic agents, lipid-lowering/lipid-modulatingagents, agents for treating diabetic complications, anti-obesity agents,antihypertensive agents, antihyperuricemic agents, and agents fortreating chronic heart failure, atherosclerosis or related disorders. Aneffective amount of the compound and/or combination partner will, ofcourse, be dependent on the subject being treated, the severity of theaffliction and the manner of administration. Determination of aneffective amount is well within the capability of those skilled in theart, especially in light of the detailed disclosure provided herein.Generally, an efficacious or effective amount of a compound isdetermined by first administering a low dose or small amount, and thenincrementally increasing the administered dose or dosages until adesired therapeutic effect is observed in the treated subject, withminimal or no toxic side effects. Applicable methods for determining anappropriate dose and dosing schedule for administration of the presentinvention are described, for example, in Goodman and Gilman's ThePharmacological Basis of Therapeutics, 11^(th) Ed., Brunton, Lazo andParker, Eds., McGraw-Hill (2006), and in Remington: The Science andPractice of Pharmacy, 21^(st) Ed., Gennaro, Ed., Lippencott Williams &Wilkins (2003), both of which are hereby incorporated herein byreference.

V. Methods of Use

The present invention further provides methods of using the crystallineforms of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolfor the prevention and treatment of disease. In one embodiment, thepresent invention provides a method of treating a disease or conditionaffected by inhibiting SGLT2, the method including administering to asubject in need thereof a therapeutically effective amount of acomposition comprising a crystalline form of the compound of the presentinvention. Diseases affected by inhibiting SGLT2 include, but are notlimited to, type 1 and type 2 diabetes mellitus, hyperglycemia, diabeticcomplications (such as retinopathy, nephropathy, neuropathy, ulcers,micro- and macroangiopathies, gout and diabetic foot disease), insulinresistance, metabolic syndrome (Syndrome X), hyperinsulinemia,hypertension, hyperuricemia, obesity, edema, dyslipidemia, chronic heartfailure, atherosclerosis, cancer and related diseases, which comprisesadministering an effective amount of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,to a subject in need thereof. In another embodiment the inventionprovides a method of using the crystalline compound, for the preparationof a medicament for treating type 1 and type 2 diabetes mellitus,hyperglycemia, diabetic complications, insulin resistance, metabolicsyndrome, hyperinsulinemia, hypertension, hyperuricemia, obesity, edema,dyslipidemia, chronic heart failure, atherosclerosis, cancer and relateddiseases. In other embodiments, the invention provides a method oftreating type 1 diabetes mellitus, type 2 diabetes mellitus,hyperglycemia, diabetic complications, insulin resistance, metabolicsyndrome, hyperinsulinemia, hypertension, hyperuricemia, obesity, edema,dyslipidemia, chronic heart failure, atherosclerosis, and cancer.

In other embodiments, the present invention provides a method oftreating diabetes, the method including administering to a subject inneed thereof a therapeutically effective amount of a compositioncomprising a crystalline form of the compound of the present invention.The diabetes can be any suitable form of diabetes, including, but notlimited to, type 1 diabetes mellitus, type 2 diabetes mellitus, anddiabetic complications. In some embodiments, the diabetes is type 1diabetes mellitus. In some other embodiments, the diabetes is type 2diabetes mellitus.

The present invention also contemplates the use of the crystalline formsof(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,in combination with other therapeutic agents, particularly those usedfor treating the above-mentioned diseases and conditions, such asantidiabetic agents, lipid-lowering/lipid-modulating agents, agents fortreating diabetic complications, anti-obesity agents, antihypertensiveagents, antihyperuricemic agents, and agents for treating chronic heartfailure, atherosclerosis or related disorders. Those skilled in the artwill appreciate that other therapeutic agents discussed below can havemultiple therapeutic uses and the listing of an agent in one particularcategory should not be construed to limit in any way its usefulness incombination therapy with compounds of the present invention.

Examples of antidiabetic agents suitable for use in combination withcrystalline forms of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolof the present invention include insulin and insulin mimetics,sulfonylureas (such as acetohexamide, carbutamide, chlorpropamide,glibenclamide, glibornuride, gliclazide, glimepiride, glipizide,gliquidone, glisoxepide, glyburide, glyclopyramide, tolazamide,tolcyclamide, tolbutamide and the like), insulin secretion enhancers(such as JTT-608, glybuzole and the like), biguanides (such asmetformin, buformin, phenformin and the like), sulfonylurea/biguanidecombinations (such as glyburide/metformin and the like), meglitinides(such as repaglinide, nateglinide, mitiglinide and the like),thiazolidinediones (such as rosiglitazone, pioglitazone, isaglitazone,netoglitazone, rivoglitazone, balaglitazone, darglitazone, CLX-0921 andthe like), thiazolidinedione/biguanide combinations (such aspioglitazone/metformin and the like), oxadiazolidinediones (such asYM440 and the like), peroxisome proliferator-activated receptor(PPAR)-gamma agonists (such as farglitazar, metaglidasen, MBX-2044, GI262570, GW1929, GW7845 and the like), PPAR-alpha/gamma dual agonists(such as muraglitazar, naveglitazar, tesaglitazar, peliglitazar,JTT-501, GW-409544, GW-501516 and the like), PPAR-alpha/gamma/delta panagonists (such as PLX204, GlaxoSmithKline 625019, GlaxoSmithKline 677954and the like), retinoid-x receptor (RXR) agonists (such as ALRT-268,AGN-4204, MX-6054, AGN-194204, LG-100754, bexarotene and the like),alpha-glucosidase inhibitors (such as acarbose, miglitol and the like),stimulants of insulin receptor tyrosine kinase (such as TER-17411,L-783281, KRX-613 and the like), tripeptidyl peptidase II inhibitors(such as UCL-1397 and the like), dipeptidyl peptidase IV inhibitors(such as sitagliptin, vildagliptin, denagliptin, saxagliptin,alogliptin, dutogliptin, NVP-DPP728, P93/01, P32/98, FE 99901, TS-021,TSL-225, GRC8200, compounds described in U.S. Pat. Nos. 6,869,947;6,727,261; 6,710,040; 6,432,969; 6,172,081; 6,011,155 and the like),glucokinase activators (such as ARRY-403, piragliatin (RO4389620),RO0281675, MK-0941, TTP355, GKA50, GKA60, GKM-001, PSN010, PSN-GK1,compounds described in Sarabu, R., et al., Expert Opinion on TherapeuticPatents, Vol. 21, No. 1, 2011, pp. 13-33, and the like), proteintyrosine phosphatase-1B inhibitors (such as KR61639, IDD-3, PTP-3848,PTP-112, OC-86839, PNU-177496, compounds described in Vats, R. K., etal., Current Science, Vol. 88, No. 2, 25 Jan. 2005, pp. 241-249, and thelike), glycogen phosphorylase inhibitors (such as N,N-4201, CP-368296and the like), glucose-6-phosphatase inhibitors, fructose1,6-bisphosphatase inhibitors (such as CS-917, MB05032 and the like),pyruvate dehydrogenase inhibitors (such as AZD-7545 and the like),imidazoline derivatives (such as BL11282 and the like), hepaticgluconeogenesis inhibitors (such as FR-225659 and the like),D-chiroinositol, glycogen synthase kinase-3 inhibitors (such ascompounds described in Vats, R. K., et al., Current Science, Vol. 88,No. 2, 25 Jan. 2005, pp. 241-249, and the like), 11 beta-hydroxysteroiddehydrogenase type 1 inhibitors (such as carbenoxolone, INCB13739 andthe like), glucagon receptor antagonists (such as BAY-27-9955, N,N-2501,NNC-92-1687 and the like), glucagon-like peptide-1 (GLP-1), GLP-1receptor agonists (such as exenatide, liraglutide, CJC-1131, AVE-0100,AZM-134, LY-315902, GlaxoSmithKline 716155 and the like), amylin, amylinanalogs and agonists (such as pramlintide and the like), fatty acidbinding protein (aP2) inhibitors (such as compounds described in U.S.Pat. Nos. 6,984,645; 6,919,323; 6,670,380; 6,649,622; 6,548,529 and thelike), beta-3 adrenergic receptor agonists (such as solabegron,CL-316243, L-771047, FR-149175 and the like), and other insulinsensitivity enhancers (such as reglixane, ONO-5816, MBX-102, CRE-1625,FK-614, CLX-0901, CRE-1633, N,N-2344, BM-13125, BM-501050, HQL-975,CLX-0900, MBX-668, MBX-675, S-15261, GW-544, AZ-242, LY-510929,AR-H049020, GW-501516 and the like).

Examples of agents for treating diabetic complications suitable for usein combination with the crystalline compound of the present inventioninclude aldose reductase inhibitors (such as epalrestat, imirestat,tolrestat, minalrestat, ponalrestat, zopolrestat, fidarestat, ascorbylgamolenate, ADN-138, BAL-ARI8, ZD-5522, ADN-311, GP-1447, IDD-598,risarestat, zenarestat, methosorbinil, AL-1567, M-16209, TAT, AD-5467,AS-3201, NZ-314, SG-210, JTT-811, lindolrestat, sorbinil and the like),inhibitors of advanced glycation end-products (AGE) formation (such aspyridoxamine, OPB-9195, ALT-946, ALT-711, pimagedine and the like), AGEbreakers (such as ALT-711 and the like), sulodexide,5-hydroxy-1-methylhydantoin, insulin-like growth factor-I,platelet-derived growth factor, platelet-derived growth factor analogs,epidermal growth factor, nerve growth factor, uridine, protein kinase Cinhibitors (such as ruboxistaurin, midostaurin and the like), sodiumchannel antagonists (such as mexiletine, oxcarbazepine and the like),nuclear factor-kappaB (NF-kappaB) inhibitors (such as dexlipotam and thelike), lipid peroxidase inhibitors (such as tirilazad mesylate and thelike), N-acetylated-alpha-linked-acid-dipeptidase inhibitors (such asGPI-5232, GPI-5693 and the like), and carnitine derivatives (such ascarnitine, levacecamine, levocarnitine, ST-261 and the like).

Examples of antihyperuricemic agents suitable for use in combinationwith the crystalline compound of the present invention include uric acidsynthesis inhibitors (such as allopurinol, oxypurinol and the like),uricosuric agents (such as probenecid, sulfinpyrazone, benzbromarone andthe like) and urinary alkalinizers (such as sodium hydrogen carbonate,potassium citrate, sodium citrate and the like).

Examples of lipid-lowering/lipid-modulating agents suitable for use incombination with the crystalline compound of the present inventioninclude hydroxymethylglutaryl coenzyme A reductase inhibitors (such asacitemate, atorvastatin, bervastatin, carvastatin, cerivastatin,colestolone, crilvastatin, dalvastatin, fluvastatin, glenvastatin,lovastatin, mevastatin, nisvastatin, pitavastatin, pravastatin,ritonavir, rosuvastatin, saquinavir, simvastatin, visastatin, SC-45355,SQ-33600, CP-83101, BB-476, L-669262, S-2468, DMP-565, U-20685,BMS-180431, BMY-21950, compounds described in U.S. Pat. Nos. 5,753,675;5,691,322; 5,506,219; 4,686,237; 4,647,576; 4,613,610; 4,499,289 and thelike), fibric acid derivatives (such as gemfibrozil, fenofibrate,bezafibrate, beclobrate, binifibrate, ciprofibrate, clinofibrate,clofibrate, etofibrate, nicofibrate, pirifibrate, ronifibrate,simfibrate, theofibrate, AHL-157 and the like), PPAR-alpha agonists(such as GlaxoSmithKline 590735 and the like), PPAR-delta agonists (suchas GlaxoSmithKline 501516 and the like), acyl-coenzyme A:cholesterolacyltransferase inhibitors (such as avasimibe, eflucimibe, eldacimibe,lecimibide, NTE-122, MCC-147, PD-132301-2, C1-1011, DUP-129, U-73482,U-76807, TS-962, RP-70676, P-06139, CP-113818, RP-73163, FR-129169,FY-038, EAB-309, KY-455, LS-3115, FR-145237, T-2591, J-104127, R-755,FCE-27677, FCE-28654, YIC-C8-434, CI-976, RP-64477, F-1394, CS-505,CL-283546, YM-17E, 447C88, YM-750, E-5324, KW-3033, HL-004 and thelike), probucol, thyroid hormone receptor agonists (such asliothyronine, levothyroxine, KB-2611, GC-1 and the like), cholesterolabsorption inhibitors (such as ezetimibe, SCH48461 and the like),lipoprotein-associated phospholipase A2 inhibitors (such as rilapladib,darapladib and the like), microsomal triglyceride transfer proteininhibitors (such as CP-346086, BMS-201038, compounds described in U.S.Pat. Nos. 5,595,872; 5,739,135; 5,712,279; 5,760,246; 5,827,875;5,885,983; 5,962,440; 6,197,798; 6,617,325; 6,821,967; 6,878,707 and thelike), low density lipoprotein receptor activators (such as LY295427,MD-700 and the like), lipoxygenase inhibitors (such as compoundsdescribed in WO 97/12615, WO 97/12613, WO 96/38144 and the like),carnitine palmitoyl-transferase inhibitors (such as etomoxir and thelike), squalene synthase inhibitors (such as YM-53601, TAK-475,SDZ-268-198, BMS-188494, A-87049, RPR-101821, ZD-9720, RPR-107393,ER-27856, compounds described in U.S. Pat. Nos. 5,712,396; 4,924,024;4,871,721 and the like), nicotinic acid derivatives (such as acipimox,nicotinic acid, ricotinamide, nicomol, niceritrol, nicorandil and thelike), bile acid sequestrants (such as colestipol, cholestyramine,colestilan, colesevelam, GT-102-279 and the like), sodium/bile acidcotransporter inhibitors (such as 264W94, S-8921, SD-5613 and the like),and cholesterol ester transfer protein inhibitors (such as torcetrapib,JTT-705, PNU-107368E, SC-795, CP-529414 and the like).

Examples of anti-obesity agents suitable for use in combination with thecrystalline compound of the present invention includeserotonin-norepinephrine reuptake inhibitors (such as sibutramine,milnacipran, mirtazapine, venlafaxine, duloxetine, desvenlafaxine andthe like), norepinephrine-dopamine reuptake inhibitors (such asradafaxine, bupropion, amineptine and the like),serotonin-norepinephrine-dopamine reuptake inhibitors (such astesofensine and the like), selective serotonin reuptake inhibitors (suchas citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine,sertraline and the like), selective norepinephrine reuptake inhibitors(such as reboxetine, atomoxetine and the like), norepinephrine releasingstimulants (such as rolipram, YM-992 and the like), anorexiants (such asamphetamine, methamphetamine, dextroamphetamine, phentermine,benzphetamine, phendimetrazine, phenmetrazine, diethylpropion, mazindol,fenfluramine, dexfenfluramine, phenylpropanolamine and the like),dopamine agonists (such as ER-230, doprexin, bromocriptine mesylate andthe like), H₃-histamine antagonists (such as impentamine, thioperamide,ciproxifan, clobenpropit, GT-2331, GT-2394, A-331440, and the like),5-HT2c receptor agonists (such as, 1-(m-chlorophenyl)piperazine (m-CPP),mirtazapine, APD-356 (lorcaserin), SCA-136 (vabicaserin), ORG-12962,ORG-37684, ORG-36262, ORG-8484, Ro-60-175, Ro-60-0332, VER-3323,VER-5593, VER-5384, VER-8775, LY-448100, WAY-161503, WAY-470,WAY-163909, MK-212, BVT.933, YM-348, IL-639, IK-264, ATH-88651, ATHX-105and the like (see, e.g., Nilsson B M, J. Med. Chem. 2006,49:4023-4034)), beta-3 adrenergic receptor agonists (such as L-796568,CGP 12177, BRL-28410, SR-58611A, ICI-198157, ZD-2079, BMS-194449,BRL-37344, CP-331679, CP-331648, CP-114271, L-750355, BMS-187413,SR-59062A, BMS-210285, LY-377604, SWR-0342SA, AZ-40140, SB-226552,D-7114, BRL-35135, FR-149175, BRL-26830A, CL-316243, AJ-9677, GW-427353,N-5984, GW-2696 and the like), cholecystokinin agonists (such asSR-146131, SSR-125180, BP-3.200, A-71623, A-71378, FPL-15849, GI-248573,GW-7178, GI-181771, GW-7854, GW-5823, and the like),antidepressant/acetylcholinesterase inhibitor combinations (such asvenlafaxine/rivastigmine, sertraline/galanthamine and the like), lipaseinhibitors (such as orlistat, ATL-962 and the like), anti-epilepticagents (such as topiramate, zonisamide and the like), leptin, leptinanalogs and leptin receptor agonists (such as LY-355101 and the like),neuropeptide Y (NPY) receptor antagonists and modulators (such asSR-120819-A, PD-160170, NGD-95-1, BIBP-3226, 1229-U-91, CGP-71683,BIBO-3304, CP-671906-01, J-115814 and the like), ciliary neurotrophicfactor (such as Axokine and the like), thyroid hormone receptor-betaagonists (such as KB-141, GC-1, GC-24, GB98/284425 and the like),cannabinoid CB1 receptor antagonists (such as rimonabant, SR147778, SLV319 and the like (see, e.g., Antel J et al., J. Med. Chem. 2006,49:4008-4016)), melanin-concentrating hormone receptor antagonists (suchas GlaxoSmithKline 803430X, GlaxoSmithKline 856464, SNAP-7941, T-226296and the like (see, e.g., Handlon A L and Zhou H, J. Med. Chem. 2006,49:4017-4022)), melanocortin-4 receptor agonists (including PT-15,Ro27-3225, THIQ, NBI 55886, NBI 56297, NBI 56453, NBI 58702, NBI 58704,MB243 and the like (see, e.g., Nargund R P et al., J. Med. Chem. 2006,49:4035-4043)), selective muscarinic receptor M₁ antagonists (such astelenzepine, pirenzepine and the like), opioid receptor antagonists(such as naltrexone, methylnaltrexone, nalmefene, naloxone, alvimopan,norbinaltorphimine, nalorphine and the like), and combinations thereof.

Examples of antihypertensive agents and agents for treating chronicheart failure, atherosclerosis or related diseases suitable for use incombination with the crystalline compound of the present inventioninclude bimoclomol, angiotensin-converting enzyme inhibitors (such ascaptopril, enalapril, fosinopril, lisinopril, perindopril, quinapril,ramipril and the like), neutral endopeptidase inhibitors (such asthiorphan, omapatrilat, MDL-100240, fasidotril, sampatrilat, GW-660511,mixanpril, SA-7060, E-4030, SLV-306, ecadotril and the like),angiotensin II receptor antagonists (such as candesartan cilexetil,eprosartan, irbesartan, losartan, olmesartan medoxomil, telmisartan,valsartan, tasosartan, enoltasosartan and the like),endothelin-converting enzyme inhibitors (such as CGS 35066, CGS 26303,CGS-31447, SM-19712 and the like), endothelin receptor antagonists (suchas tracleer, sitaxsentan, ambrisentan, L-749805, TBC-3214, BMS-182874,BQ-610, TA-0201, SB-215355, PD-180988, BMS-193884, darusentan, TBC-3711,bosentan, tezosentan, J-104132, YM-598, S-0139, SB-234551, RPR-118031A,ATZ-1993, RO-61-1790, ABT-546, enlasentan, BMS-207940 and the like),diuretic agents (such as hydrochlorothiazide, bendroflumethiazide,trichlormethiazide, indapamide, metolazone, furosemide, bumetanide,torsemide, chlorthalidone, metolazone, cyclopenthiazide,hydroflumethiazide, tripamide, mefruside, benzylhydrochlorothiazide,penflutizide, methyclothiazide, azosemide, etacrynic acid, torasemide,piretanide, meticrane, potassium canrenoate, spironolactone,triamterene, aminophylline, cicletanine, LLU-alpha, PNU-80873A,isosorbide, D-mannitol, D-sorbitol, fructose, glycerin, acetazolamide,methazolamide, FR-179544, OPC-31260, lixivaptan, conivaptan and thelike), calcium channel antagonists (such as amlodipine, bepridil,diltiazem, felodipine, isradipine, nicardipen, nimodipine, verapamil,S-verapamil, aranidipine, efonidipine, barnidipine, benidipine,manidipine, cilnidipine, nisoldipine, nitrendipine, nifedipine,nilvadipine, felodipine, pranidipine, lercanidipine, isradipine,elgodipine, azelnidipine, lacidipine, vatanidipine, lemildipine,diltiazem, clentiazem, fasudil, bepridil, gallopamil and the like),vasodilating antihypertensive agents (such as indapamide, todralazine,hydralazine, cadralazine, budralazine and the like), beta blockers (suchas acebutolol, bisoprolol, esmolol, propanolol, atenolol, labetalol,carvedilol, metoprolol and the like), sympathetic blocking agents (suchas amosulalol, terazosin, bunazosin, prazosin, doxazosin, propranolol,atenolol, metoprolol, carvedilol, nipradilol, celiprolol, nebivolol,betaxolol, pindolol, tertatolol, bevantolol, timolol, carteolol,bisoprolol, bopindolol, nipradilol, penbutolol, acebutolol, tilisolol,nadolol, urapidil, indoramin and the like), alpha-2-adrenoceptoragonists (such as clonidine, methyldopa, CHF-1035, guanabenz acetate,guanfacine, moxonidine, lofexidine, talipexole and the like), centrallyacting antihypertensive agents (such as reserpine and the like),thrombocyte aggregation inhibitors (such as warfarin, dicumarol,phenprocoumon, acenocoumarol, anisindione, phenindione, ximelagatran andthe like), and antiplatelets agents (such as aspirin, clopidogrel,ticlopidine, dipyridamole, cilostazol, ethyl icosapentate, sarpogrelate,dilazep, trapidil, beraprost and the like).

Furthermore, in another aspect, the invention provides for apharmaceutical composition comprising effective amounts of a crystallineform of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,and at least one member selected from the group of therapeutic agentslisted above as combination partners, in a pharmaceutically acceptableexcipient.

The crystalline compound of the present invention is also useful fortreatment of glucose disorders. In some embodiments, the presentinvention provides a method of decreasing blood glucose in a subject inneed thereof, the method including administering to the subject aneffective amount of a composition comprising a crystalline form of thecompound of the present invention. In other embodiments, the presentinvention provides a method of lowering plasma levels of glycatedhemoglobin (HbA1c) in a subject in need thereof, the method includingadministering to the subject an effective amount of a compositioncomprising a crystalline form of the compound of the present invention.In still other embodiments, the present invention provides a method ofincreasing the excretion of glucose in the urine of a subject in needthereof, said method comprising administering to the subject aneffective amount of a composition comprising a crystalline form of thecompound of the present invention.

The treatment of the present invention can be administeredprophylactically to prevent or delay the onset or progression of adisease or condition (such as hyperglycemia), or therapeutically toachieve a desired effect (such as a desired level of serum glucose) fora sustained period of time.

The crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triolcan be administered to a subject, e.g., a human patient, a domesticanimal such as a cat or a dog, independently or together with acombination partner, in the form of their pharmaceutically acceptablesalts or prodrugs, or in the form of a pharmaceutical composition wherethe compounds and/or combination partners are mixed with suitablecarriers or excipient(s) in a therapeutically effective amount.Consequently, the crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,and an additional active agent to be combined therewith, can be presentin a single formulation, for example a capsule or tablet, or in twoseparate formulations, which can be the same or different, for example,in the form of a kit comprising selected numbers of doses of each agent.

The appropriate dosage of compound will vary according to the chosenroute of administration and formulation of the composition, among otherfactors, such as patient response. The dosage can be increased ordecreased over time, as required by an individual patient. A patientinitially may be given a low dose, which is then increased to anefficacious dosage tolerable to the patient. Typically, a useful dosagefor adults may be from 1 to 2000 mg, preferably 1 to 200 mg, whenadministered by oral route, and from 0.1 to 100 mg, preferably 1 to 30mg, when administered by intravenous route, in each case administeredfrom 1 to 4 times per day. When a compound of the invention isadministered in combination with another therapeutic agent, a usefuldosage of the combination partner may be from 20% to 100% of thenormally recommended dose.

Dosage amount and interval can be adjusted individually to provideplasma levels of the active compounds which are sufficient to maintaintherapeutic effect. Preferably, therapeutically effective serum levelswill be achieved by administering single daily doses, but efficaciousmultiple daily dose schedules are included in the invention. In cases oflocal administration or selective uptake, the effective localconcentration of the drug may not be related to plasma concentration.One having skill in the art will be able to optimize therapeuticallyeffective local dosages without undue experimentation.

All publications and patent applications cited in this specification areherein incorporated by reference as if each individual publication orpatent application were specifically and individually indicated to beincorporated by reference. Any conflict between any reference citedherein and the teaching of this specification is to be resolved in favorof the latter. Similarly, any conflict between an art-recognizeddefinition of a word or phrase and a definition of the word or phrase asprovided in this specification is to be resolved in favor of the latter.Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it will be readily apparent to those of ordinary skill inthe art in light of the teachings of this invention that certain changesand modifications can be made thereto without departing from the spiritor scope of the appended claims. The invention will be described ingreater detail by way of specific examples.

VI. Examples

The following examples are offered for illustrative purposes, and arenot intended to limit the invention in any manner. Those of skill in theart will readily recognize a variety of noncritical parameters which canbe changed or modified to yield essentially the same results.

The names of compounds shown in the following examples were derived fromthe structures shown using the CambridgeSoft Struct=Name algorithm asimplemented in ChemDraw Ultra version 10.0. Unless otherwise indicated,the structures of compounds synthesized in the examples below wereconfirmed using the following procedures:

(1) Gas chromatography-mass spectra with electrospray ionization (MSESI) were obtained with an Agilent 5973N mass spectrometer equipped withan Agilent 6890 gas chromatograph with an HP-5 MS column (0.25 μmcoating; 30 m×0.25 mm). The ion source was maintained at 230° C. andspectra were scanned from 25-500 amu at 3.09 sec per scan.

(2) High pressure liquid chromatography mass spectra (LC-MS) wereobtained using Finnigan Surveyor HPLC equipped with a quaternary pump, avariable wavelength detector set at 254 nm, an XB-C18 column (4.6×50 mm,5 μm), and a Finnigan LCQ ion trap mass spectrometer with electrosprayionization. Spectra were scanned from 80-2000 amu using a variable iontime according to the number of ions in the source. The eluents were B:acetonitrile and D: water. Gradient elution from 10% to 90% B in 8 minat a flow rate of 1.0 mL/min is used with a final hold at 90% B of 7min. Total run time is 15 min.

(3) Routine one-dimensional NMR spectroscopy was performed on 400 MHz or300 MHz Varian Mercury-Plus spectrometers. The samples were dissolved indeuterated solvents obtained from Qingdao Tenglong Weibo Technology Co.,Ltd., and transferred to 5 mm ID NMR tubes. The spectra were acquired at293 K. The chemical shifts were recorded on the ppm scale and werereferenced to the appropriate solvent signals, such as 2.49 ppm forDMSO-d6, 1.93 ppm for CD₃CN, 3.30 ppm for CD₃OD, 5.32 ppm for CD₂Cl₂ and7.26 ppm for CDCl₃ for ¹H spectra.

When the following abbreviations and acronyms are used throughout thedisclosure, they have the following meanings: ACN, acetonitrile; Ac₂O,acetic anhydride; AcOEt, ethyl acetate; AcOH, acetic acid; A1Br₃,aluminum bromide; AlCl₃, aluminum chloride; BBr₃, boron tribromide;BF₃.Et₂O, boron trifluoride etherate; n-BuLi, n-butyllithium; s-BuLi,s-butyllithium; t-BuLi, t-butyllithium; t-BuOK, potassium tert-butoxide;CaCl₂, calcium chloride; calc., calculated; CD₃OD, methanol-d₄; CDCl₃,chloroform-d; CF₃SO₃H, trifluoromethanesulfonic acid; CH₂Cl₂, methylenechloride; CH₂I₂, methylene iodide; CH₃CN, acetonitrile; (COCl)₂, oxalylchloride; DAST, (diethylamino)sulfur trifluoride; DCM, dichloromethane;DIAD, diisopropyl azodicarboxylate; DMAP, 4-dimethylaminopyridine; DMEM,Dulbecco's Modified Eagle Medium; DMF, N,N-dimethylformamide; DMP,Dess-Martin periodinane; DMSO, dimethylsulfoxide; EA, ethyl acetate; eq,equivalents; ESI, electrospray ionization; Et, ethyl; Et₃SiH,triethylsilane; EtOAc, ethyl acetate; EtOH, ethanol; FBS, fetal bovineserum; h, hour; H₂, hydrogen gas; H₂SO₄, sulfuric acid; Hepes,4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; ¹H NMR, protonnuclear magnetic resonance; HPLC, high performance liquidchromatography; IPA, isopropyl alcohol (2-propanol); IPC, In-ProcessControl; K₂CO₃, potassium carbonate; K₂CrO₇, potassium dichromate; KOH,potassium hydroxide; LC-ESI-MS, liquid chromatography electrosprayionization mass spectrometry; LC-MS, liquid chromatography-massspectroscopy; Me, methyl; MeOH, methanol; MeSO₃H, methanesulfonic acid;Mg, magnesium; MgCl₂, magnesium chloride; min, minute; MS, massspectroscopy; MsOH, methanesulfonic acid; NaH, sodium hydride; NaHCO₃,sodium bicarbonate; NaOAc, sodium acetate; NaOH, sodium hydroxide;Na₂SO₄, sodium sulfate; NH₄Cl, ammonium chloride; Pd/C, palladium oncarbon; PE, petroleum ether; Ph, phenyl; POCl₃, phosphorus oxychloride;PPh₃, triphenylphosphine; R_(f), retention factor; rt, room temperature;SOCl₂, thionyl chloride; TBAI, tetrabutylammonium iodide; TFA,trifluoroacetic acid; THF, tetrahydrofuran; TLC, thin layerchromatography; TMS, trimethylsilyl; Tris, trishydroxymethylaminomethane(or 2-amino-2-(hydroxymethyl)propane-1,3-diol).

Example 1 Preparation of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,bis(L-proline) Complex

Example 1A Preparation of 2-cyclopropoxyethanol (1)

To a suspension of Mg powder (86.7 g, 3.6 mol) and iodine (cat) inanhydrous THF (0.7 L) was added slowly 1,2-dibromoethane (460 g, 2.4mol) in anhydrous THF (2 L) slowly at a rate as to keep the internaltemperature between 40-55° C. After the addition, a solution of2-(2-bromoethyl)-1,3-dioxolane (100 g, 0.56 mol) in anhydrous THF (750mL) was added dropwise. The reaction mixture was kept at 40-55° C. for16 h and was quenched by addition of aqueous solution of ammoniumchloride. The mixture was extracted with methylene chloride. The organiclayer was dried over sodium sulfate, and concentrated to give the titleproduct (27 g) as yellow oil, which was directly used without furtherpurification.

Example 1B Preparation of 2-cyclopropoxyethyl 4-methylbenzenesulfonate(2)

To a stirred solution of sodium hydroxide (32 g, 0.8 mol) in water (180mL) and THF (180 mL) was added Example 1A (27 g, 0.26 mol) at −5 to 0°C. Afterwards, a solution of p-toluenesulfonyl chloride (52 g, 0.27 mol)in THF (360 mL) was added dropwise. The reaction mixture was kept at −5to 0° C. for 16 h. The reaction mixture was then kept at roomtemperature for 30 min. The organic layer was separated and the aqueouslayer was extracted with ethyl acetate (2×1.0 L). The combined organiclayers were washed with brine, dried over Na₂SO₄ and concentrated to getthe crude product as yellow oil (53.3 g). It was used directly withoutfurther purification.

Example 1C Preparation of 4-(5-bromo-2-chlorobenzyl)phenol (3)

To a stirred solution of 4-bromo-1-chloro-2-(4-ethoxybenzyl)benzene (747g, 2.31 mol) in dichloromethane was added boron tribromide (1.15 kg,4.62 mol) slowly at −78° C. The reaction mixture was allowed to rise toroom temperature. When the reaction was complete as measure by TLC, thereaction was quenched with water. The mixture was extracted withdichloromethane. The organic layer was washed with aqueous solution ofsaturated sodium bicarbonate, water, brine, dried over Na₂SO₄, andconcentrated. The residue was recrystallized in petroleum ether to givethe title compound as a white solid (460 g, yield 68%). ¹H NMR (CDCl₃,400 MHz): δ 7.23˜7.29 (m, 3H), 7.08 (d, J=8.8 Hz, 2H), 6.79 (d, J=8.8Hz, 2H), 5.01 (s, 1H), 4.00 (s, 2H).

Example 1D Preparation of4-bromo-1-chloro-2-(4-(2-cyclopropoxyethoxy)benzyl)benzene (4)

A mixture of Example 1C (56.7 g, 210 mmol) and Cs₂CO₃ (135 g, 420 mmol)in DMF (350 mL) was stirred at room temperature for 0.5 h. Example 1B(53.3 g, 210 mmol) was added. The reaction mixture was stirred at roomtemperature overnight. It was diluted with water (3 L) and extractedwith EtOAc. The organic layer was washed with water, brine, dried overNa₂SO₄, and concentrated. The residue was purified by flash columnchromatography on silica gel eluting with petroleum ether:ethyl acetate(10:1) to give the title compound as liquid (51 g, yield 64%). ¹H NMR(CDCl₃, 400 MHz): δ 7.22˜7.29 (m, 3H), 7.08 (d, J=8.8 Hz, 2H), 6.88 (d,J=8.8 Hz, 2H), 4.10 (t, J=4.8 Hz, 2H), 3.86 (t, J=4.8 Hz, 2H), 3.38-3.32(m, 1H), 0.62-0.66 (m, 2H), 0.49-0.52 (m, 2H).

Example 1E Preparation of(2S,3R,4S,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3,4,5-triol(5)

To a stirred solution of Example 1D (213 g) in anhydrous THF/toluene(1:2 (v/v), 1.7 L) under argon was added n-BuLi (2.5 M hexane, 245.9 mL)drop wise at −60±5° C. The mixture was stirred for 30 min. beforetransferred to a stirred solution of2,3,4,6-tetra-O-trimethylsilyl-β-D-glucolactone (310.5 g) in toluene(1.6 L) at −60±5° C. The reaction mixture was continuously stirred at−60±5° C. for 1 h before quenching with aqueous solution of saturatedammonium chloride (1.5 L). Then mixture was allowed to warm to roomtemperature and stirred for 1 h. The organic layer was separated and thewater layer was extracted with ethyl acetate (3×500 mL). The combinedorganic layers were washed with brine (1 L), dried over Na₂SO₄, andconcentrated. The residue was dissolved in methanol (450 mL) andmethanesulfonic acid (9.2 mL) was added at 0° C. The solution wasallowed to warm to room temperature and stirred for 20 h. It wasquenched with aqueous solution of sodium bicarbonate (50 g) in water(500 mL) and additional water (900 mL) was added. The mixture wasextracted with ethyl acetate (3×1.0 L). The combined organic layers werewashed with brine, dried over Na₂SO₄, concentrated and used directly inthe next step without further purification.

Example 1F Preparation of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol,bis(L-proline) Complex (7)

To stirred solution of Example 1E in CH₂Cl₂/CH₃CN (650 mL:650 mL) at −5°C. was added triethylsilane (28.2 mL, 563 mmol), and followed byBF₃.Et₂O (52.3 mL, 418.9 mmol). The reaction was stirred for 16 h whilethe temperature was allowed to warm to room temperature gradually. Thereaction was quenched with aqueous solution of saturated sodiumbicarbonate to pH 8.0. The organic volatiles were removed under vacuum.The residue was partitioned between ethyl acetate (2.25 L) and water(2.25 L). The organic layer was separated, washed with brine, dried overNa₂SO₄ and concentrated to give the crude product 6 (230 g, purity82.3%). This product and L-proline (113.7 g) in EtOH/H₂O (15:1 v/v, 2.09L) was stirred at 80° C. for 1 h when it became a clear solution. Hexane(3.0 L) was added dropwise into the above hot solution over 50 min, withthe temperature being kept at about 60° C. The reaction mixture wasstirred overnight at room temperature. The solid was filtered and washedwith EtOH/H₂O (15:1 (v/v), 2×300 mL), hexane (2×900 mL), and dried at45° C. under vacuum for 10 h to give the pure title compound 7 as awhite solid (209 g). Purity (HPLC) 99.2% (UV). ¹H NMR (CD₃OD, 400 MHz):δ 7.25˜7.34 (m, 3H), 7.11 (d, J=8.8 Hz, 2H), 6.84 (d, J=8.8 Hz, 2H),4.03-4.11 (m, 5H), 3.96-4.00 (m, 2H), 3.83-3.90 (m, 3H), 3.68-3.72 (m,1H), 3.36-3.46 (m, 6H), 3.21-3.30 (m, 3H), 2.26-2.34 (m, 2H), 2.08-2.17(m, 2H), 1.94-2.02 (m, 4H), 0.56-0.57 (m, 2H), 0.52-0.53 (m, 2H).

Example 2 Direct Preparation of Crystalline Compound 8 from Complex 7

This example illustrates the preparation of a crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

To a 5.0 L 4-necked flask equipped with a mechanical stirrer was addedthe starting co-crystal (150.0 g) and methanol (300 mL). The mixture wasstirred at room temperature with mechanical stirring (anchor agitator,2-blades 9 cm) until a cloudy solution/suspension formed, to whichdistilled water (1500 mL) was added dropwise at a rate of ˜12.5 mL/min.As the mixture warmed from the exotherm of adding water to methanol, themixture became clear after adding about ⅕ to ⅓ of the water. After theaddition was completed the reaction was stirred continuously at 80 rpmfor another 5 h. The reaction mixture was filtered over medium-speedfilter paper and the filter cake was washed with distilled water (450 mLand then 300 mL) and dried under vacuum using an oil pump (˜6 mm Hg) at45° C. for 48 hours to give the target product as a white crystallinesolid (94.2 g, 93.9% yield, purity (HPLC): 99.3%).

Example 3 Direct Preparation of Crystalline Compound 8 from Complex 7

This example illustrates alternative conditions for preparingcrystalline(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol

Procedure A:

A 250 mL of 4-neck flask was charged with the starting complex (10.0 g)and methanol (33.5 mL). After refluxing for 20 min with mechanicalstirring a clear solution formed. Water (67.0 mL) was added slowlydropwise to it over 20 min. The reaction mixture was cooled slowly toroom temperature (25° C.) in oil bath and stirred for another 3 h atroom temperature. The reaction mixture was filtered by filter paper andthe filter cake was washed with water (2×20 mL), dried under vacuum at65° C. for 8 h to give a white crystalline solid. Yield: 6.0 g (89.6%)

Procedure B:

A 250 mL of 4-neck flask was charged with the starting complex (10.0 g)and methanol (33.5 mL). After stirring for 20 min with mechanicalstirring, the solids did not completely dissolve. Water (67.0 mL) wasadded slowly dropwise to it over 20 min. At first all the remainingsolids dissolved and later new crystals started to form. The reactionmixture was stirred for another 3 h at room temperature. The reactionmixture was filtered over filter paper and the filter cake was washedwith water (2×20 mL), dried under vacuum at 65° C. for 8 h to give awhite crystalline solid. Yield: 6.0 g (89.6%).

Procedures A and B are summarized in the table below with otherconditions for preparing crystalline 8 directly from complex 7.

TABLE 1 Summary Table of Crystallization Conditions Complex (g) Methanol(mL) Water (mL) Temp. (° C.) Yield (%) 4.0 20.0 80.0 70 87.4 10.0 33.567.0 25 89.6 10.0 33.5 100.0 25 91.1 10.0 33.5 67.0 70 89.6

Example 4 Direct Preparation of Crystalline Compound 8 from Complex 7

This example illustrates the preparation of a crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

Compound 7 (14.0 kg) was dissolved in methanol (36.2 kg) and deionized(DI) water (11.2 kg) and then filtered. Additional DI water (41.3 kg)was added and then seed crystals were added at 35±5° C. to crystallizecompound 8 from solution. Additional DI water (41.3 kg) was added tocomplete precipitation. The resulting slurry was filtered, and theproduct solids rinsed on the filter with DI water, transferred to trays,and dried under vacuum at ˜65° C. to afford 8.75 kg of compound 8.

Example 5 Indirect Preparation of Crystalline Compound 8 from Complex 7

To a 200 L glass lined reactor equipped with a double-tier paddleagitator and a glass condenser was added sequentially complex 7 (7.33kg), ethyl acetate (67.5 kg) and pure water (74.0 kg). The mixture washeated to reflux and stirred at reflux for 30 min. The reaction mixturewas cooled to approximately 50° C. and the organic layer was separatedand the aqueous layer was extracted with ethyl acetate (34.0 kg). Thecombined organic layers were washed with pure water (3×74.0 kg) (IPCtest showed that the IPC criteria for L-proline residue was met afterthree water washes). The mixture was concentrated at 40° C. under vacuum(˜15 mmHg) for 3 h until the liquid level dropped below the lower-tieragitator paddle. The mixture (18 kg) was discharged and transferred to a20 L rotary evaporator. The mixture was concentrated under vacuum (40°C., ˜5 mmHg) to a minimum volume. The remaining trace amount of ethylacetate was removed azeotropically at 40° C. under vacuum with methanol(10 kg). The residue was dried under vacuum of an oil pump (˜6 mmHg) at40° C. for 10 h to give 8 as a white amorphous solid (4.67 kg, purity(HPLC): 99.2%) which was used in the next step without furtherpurification.

The recrystallization was accomplished by the following steps. To a 100L glass line reactor equipped with a double-tier paddle agitator and aglass condenser was added the above amorphous 8 (4.67 kg) and methanol(18.0 kg). The mixture was refluxed at 70° C. for 30 min until a clearsolution formed, to which pure water (45.0 kg) was added over 2 hours.After the addition was completed (the reaction temperature was 41° C.),the reaction mixture was cooled to room temperature and stirred at roomtemperature for 15 hours. The reaction mixture was filtered and the wetcake was washed with pure water (2×15 kg) and dried under vacuum at55˜60° C. for 12 hours to give the target product as an off-whitecrystalline solid (3.93 kg, yield: 84% in two steps; purity (HPLC):99.7%).

Example 6 Direct Preparation of Crystalline Compound 8 from Amorphous 8

5 L 4-neck flask was charged with 8 (amorphous), 116 g, and methanol(580 mL). The reaction mixture was heated to 60 C with mechanicalstirring and the solution became clear. Water (2320 mL) was addeddropwise to the reaction solution at 40 mL/min at 50° C. The reactionmixture was stirred overnight at room temperature. The reaction mixturewas filtered and the filter cake was washed with water (2×200 mL), driedunder vacuum at 55° C. for 12 hours, to afford white crystalline 8.Yield is 112.8 g (97.2%).

Example 7 Direct Preparation of Crystalline Compound 8 from Crude 6(with Seed Crystal)

This example illustrates the preparation of a crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

A 250 mL 4-neck flask was charged with 6 (12.0 g, HPLC Purity: 88.3%)and methanol (48 mL). After refluxing for 30 min with magnetic stirring(120 RPM), water (72 mL) was added dropwise to the above solution over20 min. After refluxing for another 30 min, the mixture was slowlycooled to 40 to 45° C., and seed crystal (10 mg) was added. Afterstirring for another 2 hours at 35 to 40° C., the mixture was cooledslowly to 20 to 25° C. and stirred for another 16 hours. The mixture wasfiltered and the filter cake was washed with water (2×24 mL), driedunder vacuum at 60 to 65° C. for 12 hours, to afford off-whitecrystalline 8. Yield is 10.6 g (88.3%). HPLC Purity: 91.8%.

Example 8 Direct Preparation of Crystalline Compound 8 from Crude 6(without Seed Crystal)

This example illustrates the preparation of a crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

A 100 mL 3-neck flask was charged with 6 (5.0 g, HPLC Purity: 90.7%) andmethanol (20 mL). After refluxing for 30 min with magnetic stirring (120RPM), water (30 mL) was added dropwise to the above solution over 20min. After refluxing for another 30 min, the mixture was slowly cooledto 20 to 25° C. over 3 hours. After stirring for another 60 hours at 20to 25° C., the mixture was filtered and the filter cake was washed withwater (2×10 mL), dried under vacuum at 60 to 65° C. for 12 hours, toafford off-white crystalline 8. Yield is 4.3 g (86%). HPLC Purity:92.6%.

Example 9 Preparation of Crystalline Compound 8 by Single Solvent

This example illustrates the preparation of a crystalline form of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.

A 40 mL glass bottle was charged with(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol(300 mg, HPLC Purity: 99.6%) and ethanol (10 mL). After shaking for 15min at 20 to 25° C., the solid was absolutely dissolved. The solutionwas kept quiescence but allowed the solvent to evaporate slowly. After 2weeks, there were only about 2 mL ethanol left and lots of needlecrystals were formed. The mixture was filtered, dried under vacuum at 60to 65° C. for 12 hours, to afford white crystalline 8. Yield is 246 mg(82%). HPLC Purity: 99.7%.

Example 10 Recrystallization of Compound 8

About 100 mg of crystalline compound 8 was dissolved with a minimalamount of solvent at about 60° C. The solution was filtrated andseparated into two parts, with one part cooled in an ice bath andagitated (quick), and the other allowed to cool naturally by exposure toambient atmosphere and temperature (slow). The solids were collected ona filter, dried and analyzed by XRPD. The table below summarizes thesolvents and results of the recrystallization. All crystals formed areidentical to the XRPD of the starting material.

TABLE 2 Summary of Crystallization of Compound 8 from Hot SaturatedSolutions Solvents Methods¹ Results² ACN quick Crystal, no change slowCrystal, no change 95% EtOH quick Crystal, no change slow Crystal, nochange EtOAc quick Crystal, no change slow Crystal, no change IPA quickCrystal, no change slow Crystal, no change Butanol quick Crystal, nochange slow Crystal, no change Butanone quick Crystal, no change slowCrystal, no change 1,4-dioxane-Heptane (1:1) quick Crystal, no changeslow No crystal IPA-EtOAc (1:1) quick Crystal, no change slow Crystal,no change ¹Quick = cooling in an ice bath; slow = cooling by exposure toambient atmosphere and temperature. ²No change = XRPD of productidentical to XRPD of starting material.

Example 11 Low Hygroscopicity of Crystalline Compound 8

The tendency toward hygroscopicity of crystalline compound 8 in powderform was tested at 75% and 92.5% relative humidity at 25° C. for up to10 days. Reagents included: (1) water: in house, MilliQ, 18.2MΩ; (2)NaCl: AR grade; (3) KNO₃: AR grade; (4) saturated NaCl solution withextra NaCl solid for 25° C./75% RH control; and (5) saturated KNO₃solution with extra KNO₃ solid for 25° C./92.5% RH control. Equipmentused included (1) desiccators, 240 mm ID; and (2) weighing bottles withlids: 50 mm ID×30 mm height.

The saturated salt solutions and solids were transferred into individualdesiccators, and equilibrated at 25° C. at least overnight to reach thedesired relative humidity readings. Four weighing bottles were placedinto each desiccator, and equilibrated overnight. The empty weighingbottles were weighed and the tare weights recorded (W₁). 0.5 g ofcrystalline compound 8 was added into three weighing bottles in eachdesiccator to form a thin layer with thickness of 1-2 mm. The sampleweight in each bottle (W₂) was recorded. One empty weighing bottle wasused for blank calibration. The bottles were left in each desiccatorwith lids open. The lids were closed and each bottle accurately weighedon days 1, 5 and 10 (W₃). The bottles were returned to each desiccatorwith lids open immediately after weighing.

The following formula was used to calculate the weight increase:

${{Weight}\mspace{14mu} {Increase}\mspace{14mu} \%} = {\frac{W_{3} - W_{1} - W_{2} - W_{B}}{W_{2}} \times 100\%}$W_(B) = the  weight  increase  of  empty  weighing  bottle.

TABLE 3 Test results of crystal compound 8 at 25° C./75% RH. EmptySample Bottle + Sample Weight Wt bottle Weight Appearance Sample WeightIncrease Appearance Initial 1 day 1 30.36142 0.50434 White powder30.86727 0.50585 0.32914% White powder 2 32.96588 0.50631 White powder33.47323 0.50735 0.23503% White powder 3 31.27798 0.50066 White powder31.77934 0.50136 0.16978% White powder Blank 31.92783 31.92768 −0.00015Average 0.24465% 5 days 1 30.86648 0.50506 0.10509% White powder 233.47339 0.50751 0.19948% White powder 3 31.77962 0.50164 0.15779% Whitepowder Blank 31.92802 0.00019 Average 0.15412% 10 days 1 30.867380.50596 0.35095% White powder 2 33.47321 0.50733 0.23108% White powder 331.77947 0.50149 0.19574% White powder Blank 31.92768 −0.00015 Average0.25926%

TABLE 4 Test results of crystal compound 8 at 5° C./92.5% RH. EmptySample Bottle + Sample Weight bottle Weight Appearance Sample WeightIncrease Appearance Initial 1 day 1 34.11948 0.50356 White powder34.62330 0.50382 0.05362% White powder 2 30.13094 0.50215 White powder30.63360 0.50266 0.10355% White powder 3 33.01277 0.50546 White powder33.51923 0.50646 0.19982% White powder Blank 40.35822 40.35821 −0.00001Average 0.11900% 5 days 1 34.62358 0.50410 0.04170% White powder 230.63446 0.50352 0.20711% White powder 3 33.51977 0.50700 0.23939% Whitepowder Blank 40.35855 0.00033 Average 0.16273% 10 days 1 34.624720.50524 0.26015% White powder 2 30.63428 0.50334 0.16330% White powder 333.51994 0.50717 0.26511% White powder Blank 40.35859 0.00037 Average0.22952%

The weight increases of crystalline compound 8 in powder samples at 75%and 92.5% relative humidity at 25° C. for 10 days were below 0.26%.Thus, crystalline compound 8 in powder form exhibited low hygroscopicityat the study conditions.

Example 12 Preparation of Capsules Containing Crystalline Compound 8

To prepare capsules containing crystalline compound 8, the compound andsilicified microcrystalline cellulose (Prosolv HD90) were blended andthen sifted through a #30 sieve into a polyethylene bag. A portion ofthe crystalline compound 8/Prosolv HD90 blend was removed. Magnesiumstearate was sifted through a #30 sieve into this portion and themixture blended. The 3-component portion was returned to the largercompound 8/Prosolv HD90 blend, and the mixture further blended. Thefinal blend was put into a polyethylene bag. Empty capsule shells (Size2) were weighed to determine the average capsule weight. The final blendin the polyethylene bag was fed onto an MG2 Planeta encapsulator andapproximately 100 mg of final blend was loaded into each capsule. Aboutevery 5 to 10 minutes, loaded capsules were sampled for acceptable fillweight. This was achieved by sampling 10 loaded capsules, weighing eachloaded capsule and comparing the results to the theoretical averageweight of a capsule plus the targeted 100 mg load of blend. Another 10capsules were visually inspected for cracks, chips, dents, splitsunexpected marks, and closures. If necessary, the encapsulation andblend loading process can be adjusted to maintain proper target weightfill into the capsules. Acceptable capsules were placed in a doublelined polyethylene bag for further processing. Loaded capsules wereweight sorted electronically, with low and high weigh filled capsulesrejected, and acceptable weight filled capsules forwarded for furtherprocessing. Acceptable weight checked capsules were polished and placedin double polyethylene bags.

TABLE 5 Components of 20 mg Capsules with Crystalline 8 Amount per unitQuality Component (mg/capsule) Function Standard Crystalline Compound 820.00 Active In house Ingredient Silicified Microcrystalline 79.40Glidant and USP/NF; Cellulose (Prosolv ® HD90) Hydrophilic Ph. Eur.,Matrix JP. Magnesium Stearate 0.60 Lubricant USP/NF; (HyQual ®,Vegetable Ph. Eur., Grade) JP Size 2, Gelatin, White, 1 Capsule Shell Inhouse Coni-Snap ® (0999) Opaque Body and Cap

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, one of skill in the art will appreciate that certainchanges and modifications may be practiced within the scope of theappended claims. In addition, each reference provided herein isincorporated by reference in its entirety to the same extent as if eachreference was individually incorporated by reference. Where a conflictexists between the instant application and a reference provided herein,the instant application shall dominate.

1. A crystalline form of the compound of the formula:


2. The crystalline form in accordance with claim 1, characterized by an X-ray powder diffraction (XRPD) pattern that comprises one or more peaks at 5.4, 11.2, 11.3, 11.9, 12.9, 15.5, 16.3, 17.8, 19.1, 20.0, 20.6, 20.7, 21.2, 22.8, 23.0, 23.4, 23.6, 23.9, 24.7, 25.4, 25.8, 27.8 and 28.2 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation.
 3. The crystalline form in accordance with claim 2, characterized by an XRPD that comprises peaks at 12.9, 19.1 and 20.7 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation.
 4. The crystalline form in accordance with claim 2, characterized by an XRPD that comprises peaks at 11.2, 12.9, 15.5, 17.8, 19.1, 20.0 and 20.7 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation.
 5. The crystalline form in accordance with claim 2, characterized by an XRPD that comprises peaks at 11.2, 12.9, 15.5, 17.8, 19.1, 20.0, 20.6, 20.7, 21.2 and 22.8 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation.
 6. The crystalline form in accordance with claim 2, characterized by an XRPD that comprises peaks at 5.4, 11.2, 11.9, 12.9, 15.5, 16.3, 17.8, and 19.1 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation.
 7. The crystalline form in accordance with claim 2, characterized by an X-ray powder diffraction (XRPD) pattern that comprises peaks at 11.2 and 12.9 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation.
 8. The crystalline form in accordance with claim 1, characterized by the XRPD peaks substantially in accordance with those of FIG.
 2. 9. The crystalline form in accordance with claim 1, characterized by a Raman spectra that comprises one or more peaks at about 353, 688, 825, 1178, 1205, 1212, 1608, 2945, 3010 and 3063 cm⁻¹.
 10. The crystalline form in accordance with claim 1, characterized by a Raman spectra that comprises peaks at about 353, 688, and 825 cm⁻¹.
 11. The crystalline form in accordance with claim 1, characterized by the Raman peaks substantially in accordance with those of FIG.
 4. 12. The crystalline form in accordance with claim 1, characterized by an X-ray powder diffraction (XRPD) pattern that comprises one or more peaks at 5.4, 11.2, 11.3, 11.9, 12.9, 15.5, 16.3, 17.8, 19.1, 20.0, 20.6, 20.7, 21.2, 22.8, 23.0, 23.4, 23.6, 23.9, 24.7, 25.4, 25.8, 27.8 and 28.2 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation; and a Raman spectra that comprises one or more peaks at about 353, 688, 825, 1178, 1205, 1212, 1608, 2945, 3010 and 3063 cm⁻¹.
 13. The crystalline form in accordance with claim 12, characterized by an X-ray powder diffraction (XRPD) pattern that comprises one or more peaks at 11.2, 12.9, 15.5, 17.8, 19.1, 20.0, 20.6, 20.7, 21.2 and 22.8 and 28.2 degrees 2θ (±0.1 degrees 2θ), wherein said XRPD is made using CuK_(α1) radiation; and a Raman spectra that comprises one or more peaks at about 353, 688, and 825 cm⁻¹.
 14. The crystalline form in accordance with claim 1, characterized by a DSC endotherm at about 136° C.
 15. The crystalline form in accordance with claim 1, characterized by the unit cell data substantially in accordance with FIG.
 7. 16. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a crystalline form of the compound (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.
 17. A crystalline form of a compound of claim 1 which is isotopically labeled.
 18. A method for making a crystalline form of the compound of claim 1, said method comprising the steps: (a) combining (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol bis(L-proline) complex and a suitable solvent with mixing to form a solution; (b) adding a precipitating solvent to said solution to provide a mixture; and (c) isolating said crystalline form from said mixture.
 19. The method of claim 18, wherein the solvent of step (a) is selected from the group consisting of methanol and ethanol.
 20. The method of claim 18, wherein the solvent of step (a) is methanol.
 21. The method of claim 18, wherein the precipitating solvent is water.
 22. The method of claim 18, wherein the solvent of step (a) is methanol and the precipitating solvent is water.
 23. The method of claim 22, wherein the ratio of methanol to water in the mixture of step (b) is from about 1:1 to about 1:9 by volume.
 24. The method of claim 22, wherein the ratio of methanol to water in the mixture of step (b) is about 1:5 by volume.
 25. The method of claim 18, wherein the ratio of complex to solvent and precipitating solvent in the mixture of step (b) is from about 1:10 to about 1:25 (w/v).
 26. The method of claim 18, wherein the mixture of step (b) further comprises a base.
 27. The method of claim 18, wherein the mixture of step (b) further comprises a salt.
 28. The method of claim 18, wherein the mixture of step (b) further comprises a seed crystal of the compound of claim
 1. 29. A method for making a crystalline form of the compound of claim 1, said method comprising the steps: (a) combining (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol and a suitable solvent with mixing to form a solution; and (b) isolating said crystalline form from said solution.
 30. The method of claim 29, said method comprising the steps: (a) combining amorphous (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol and a suitable solvent with mixing to form a solution; and (b) adding a precipitating solvent to said solution to form a mixture; and (c) isolating said crystalline form from said mixture.
 31. The method of claim 29, further comprising adding a precipitating solvent to said solution.
 32. The method of claim 31, wherein the solvent of step (a) is selected from the group consisting of methanol and ethanol.
 33. The method of claim 31, wherein the solvent of step (a) is methanol.
 34. The method of claim 31, wherein the precipitating solvent is water.
 35. The method of claim 31, wherein the solvent of step (a) is methanol and the precipitating solvent is water.
 36. The method of claim 35, wherein the ratio of methanol to water in the mixture of step (b) is from about 1:1 to about 1:9 by volume.
 37. The method of claim 35, wherein the ratio of methanol to water in the mixture of step (b) is about 1:5 by volume.
 38. The method of claim 31, wherein the ratio of complex to solvent and precipitating solvent in the mixture of step (b) is from about 1:1 to about 1:9 by volume.
 39. The method of claim 31, wherein the solution further comprises a seed crystal of the compound of claim
 1. 40. The method of claim 31, wherein the (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol is amorphous (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol.
 41. The method of claim 40, wherein the amorphous (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol is prepared from (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol bis(L-proline) by (a2) combining (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol bis(L-proline) complex and a suitable solvent mixture with mixing to form a solution; and (a3) isolating amorphous (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-(2-cyclopropoxyethoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol from said solution of step (a2).
 42. The method of claim 41, wherein said solvent mixture of step (a2) comprises an organic solvent and an aqueous solvent.
 43. The method of claim 41, wherein said solvent mixture of step (a2) comprises ethyl acetate and water.
 44. A method of treating a disease or condition affected by inhibiting SGLT2, said method comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a crystalline form of the compound of claim 1, wherein said disease or condition is selected from the group consisting of type 1 diabetes mellitus, type 2 diabetes mellitus, hyperglycemia, diabetic complications, insulin resistance, metabolic syndrome, hyperinsulinemia, hypertension, hyperuricemia, obesity, edema, dyslipidemia, chronic heart failure, atherosclerosis, and cancer.
 45. A method of treating a disease or condition affected by inhibiting SGLT2, said method comprising administering to a subject in need thereof a therapeutically effective amount of a composition comprising a crystalline form of the compound of claim
 1. 